Ion Hi-Q View Chemistry for Ion Torrent Next-Generation Sequencing

The new Ion Hi-Q™ View chemistry for research using the Ion PGM™ , Ion Chef™, and Ion OneTouch™ 2 systems takes everything that made the previous Hi-Q chemistry robust and reliable, and advances it to a new level. In addition to high sequencing accuracy, Ion Hi-Q View chemistry enables enhanced experimental reproducibility and sequence coverage uniformity, helping you to answer questions with even greater confidence.

Our most advanced sequencing chemistry for the Ion PGM System

Experience the superior speed of the Ion PGM System, now with our most accurate and robust sequencing chemistry. The original Ion Hi-Q chemistry reduced variant false positives and indel errors and increased read lengths and throughput, helping to improve sequencing performance (see the data). The new Ion Hi-Q View kit is designed to deliver powerful enhancements to the previous Ion Hi-Q chemistry for improved variant detection accuracy, to help you achieve even greater confidence in your results at no additional cost. With this latest sequencing chemistry, we observed significant improvement in the total number of quality bases for targeted sequencing (total AQ20 bases) when compared to the original Hi-Q chemistry (Figure 1). The improved total quality bases per targeted sequencing run were due to better end-to-end amplicon coverage, uniformity, and the number of amplicons with no strand bias (Figure 2).

The new Ion Hi-Q View kit also helps to enhance reproducibility by minimizing user error in manual chip-loading, with visible Ion Sphere™ Particles (ISPs) that can be easily visualized during sequencing template preparation using the Ion OneTouch 2 System.

Figure 2.Improvements to targeted sequencing. Comparison of end-to-end amplicon coverage, uniformity, and number of amplicons with reduced strand bias using a custom Ion AmpliSeq panel targeting ~48,000 human exonic amplicons. Amplicons were more efficiently sequenced using the new Ion Hi-Q View Chef sequencing kit (**N=11) when compared to the previous Ion Hi-Q Chef sequencing kit (*N=8).
N=number of sequencing run

Accurate sequencing for robust variant detection

Ion Hi-Q View chemistry was designed to deliver sequencing accuracy that is equivalent to or better than that of the previous Ion Hi-Q chemistry for the Ion PGM System, while maintaining the same high-quality variant calling. Ion Hi-Q View Chef chemistry, when used for targeted sequencing of a custom ~48,000-amplicon Ion AmpliSeq panel, demonstrated improved single nucleotide variant (SNV) sensitivity (Figure 3A) compared to the previous Ion Hi-Q chemistry, while decreasing the false positive rate for indel variants (Figure 3B).

Hi-Q chemistry and Hi-Q View chemistry improve sequencing accuracy for the Ion PGM System (Figure 4), resulting in high-quality variant calling accuracy using just 10 ng of DNA input compared to the 150 ng recommended for the competitor. It also helps improve de novo assembly with up to a 90% decrease in indel error rates compared to previous Ion PGM System sequencing chemistries (Figure 5), making it ideal for microbial sequencing applications.

The Ion PI™ Hi-Q™ kit is available for use with the Ion Proton™ System to enable improved variant calling performance compared to our previous chemistries. The Hi-Q chemistry helps reduce variant false positive rates by up to 71% for exome sequencing (Figure 6), using the Ion Proton System.

Reduced burden of orthogonal confirmation

“We’ve always been happy with the Ion AmpliSeq™ Exome data but with the new Ion PI™ Hi-Q™ chemistry we get even better data. We’ve noticed a 50% reduction in indel false positives, which will give us a shorter candidate list, making it ideal for de novo or trio experiments.”Adam Ameur, Bioinformatician, Genome Center, Uppsala University